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Eur J Cardiothorac Surg 2005;27:104-110
© 2005 Elsevier Science NL

Morphological, electrophysiological and coupling characteristics of bone marrow-derived mononuclear cells—an in vitro-model

Department of Cardiac Surgery, Heart Center Leipzig, University of Leipzig, Leipzig, Germany

Received 9 February 2004; received in revised form 16 August 2004; accepted 23 August 2004.

^* Corresponding author. Tel.: +49 341 865 1421; fax: +49 341 869 6646. (E-mail: rastan{at}rz.uni-leipzig.de).

Objective: Transplantation of bone marrow-derived mononuclear cells (BMC) may be suitable to prevent myocardial remodeling and improve left ventricular function after myocardial infarction. However, it is unknown whether or not cardiomyocytes and BMCs can form functioning cell-to-cell coupling and develop adequate electrophysiological properties. Methods: BMCs were isolated from minipig leg bones, treated with 5-azacytidine (10µM) for 24h, cultured for 7 days and labeled with a fluoroscopic dye (DIL). BMCs were cocultured with spontaneously beating cardiomyocytes of neonatal rats. On days 4, 7 and 14 cocultured cells were analyzed. Immunhistochemistry (Connexin 43, -actinin) was used to assess cardiomyogenic differentiation. Action potential characteristics were recorded in whole cell patch clamp mode and to investigate intercellular communication a second gap junction permeable fluoroscopic dye was brought into BMCs by microinjection (Lucifer yellow, LY). Results: From day 7 in coculture BMCs beated synchronously with neonatal rat cardiomyocytes. On day 14, 55.9% of BMCs expressed -actinin and 98.3% were positive for gap junction protein connexin 43. BMC action potential duration (APD₉₀) was mean 11.1ms with dV/dt_max of 26.8V/s and similar to atrial cardiac type. However, microinjection of LY revealed only little dye transfer into adjacent rat cardiomyocytes. Conclusions: Cocultured BMCs have the potential for early expression of muscle specific proteins in about 60% after 14 days and for cardiac gap junction proteins. Synchronous beating indicates an effective electromechanical coupling. In this heterologous setting we could prove only weak metabolic coupling.

Key Words: Bone marrow cells • Cell transplantation • Action potential • Patch clamp • Transdifferentiation

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